title: Cell to aperture interaction in patch-clamp chips visualized by fluorescence microscopy and focused-ion beam sections.
abstract: Patch-clamp is an important method to monitor the electrophysiological activity of cells and the role of pharmacological compounds on specific ion channel proteins. In recent years, planar patch-clamp chips have been developed as a higher throughput approach to the established glass-pipette method. However, proper conditions to optimize the high resistance cell-to-probe seals required to measure the small currents resulting from ion channel activity are still the subject of conjecture. Here, we report on the design of multiple-aperture (sieve) chips to rapidly facilitate assessment of cell-to-aperture interactions in statistically significant numbers. We propose a method to pre-screen the quality of seals based on a dye loading protocol through apertures in the chip and subsequent evaluation with fluorescence confocal microscopy. We also show the first scanning electron micrograph of a focused ion beam section of a cell in a patch-clamp chip aperture.
authors.affiliation: Institute for Microstructural Sciences, National Research Council of Canada, 1200 Montreal Rd, Ottawa, Ontario K1A0R6, Canada. christophe.py@nrc.ca
authors.forename: Christophe
authors.initials: C
authors.lastname: Py
authors.forename: Danish
authors.initials: D
authors.lastname: Salim
authors.forename: Robert
authors.initials: R
authors.lastname: Monette
authors.forename: Tanya
authors.initials: T
authors.lastname: Comas
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Fraser
authors.forename: Dolores
authors.initials: D
authors.lastname: Martinez
authors.forename: Marzia
authors.initials: M
authors.lastname: Martina
authors.forename: Geoffrey
authors.initials: G
authors.lastname: Mealing
mesh_terms: D000818:Animals; D002460:Cell Line; D002477:Cells / Q000187:drug effects*; D057166:High-Throughput Screening Assays / Q000379:methods; D007473:Ion Channels / Q000378:metabolism*; D051379:Mice; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D018408:Patch-Clamp Techniques
chemical_list: D007473:Ion Channels